中华皮肤科杂志 ›› 2010, Vol. 43 ›› Issue (8): 534-537.

• 论著 • 上一篇    下一篇

rDNA-RFLP多酶切技术用于人类致病相关毛孢子菌种间鉴定初探

吕雪莲1,张晓利2,王淼淼3,刘泽虎4,沈永年2,刘维达2   

  1. 1. 大连市皮肤病医院
    2. 南京 中国医学科学院北京协和医学院皮肤病研究所
    3. 中国医学科学院皮肤病研究所
    4. 中国医学科学院、中国协和医科大学皮肤病研究所
  • 收稿日期:2010-01-07 修回日期:2010-03-22 出版日期:2010-08-15 发布日期:2010-08-10
  • 通讯作者: 吕雪莲 E-mail:snowlotus.lv@gmail.com
  • 基金资助:

    科技重大专项;卫生部部属（管）医院临床学科重点项目;江苏省自然科学基金;江苏省卫生厅基金;2006杨森JRC基金

Primary exploration on identification of pathogenic Trichosporon spp. with rDNA-RFLP analysis

  • Received:2010-01-07 Revised:2010-03-22 Online:2010-08-15 Published:2010-08-10
  • Contact: LU Xue-lian E-mail:snowlotus.lv@gmail.com

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摘要：

目的 探讨rDNA-ITS/IGS1-RFLP多酶切技术构建毛孢子菌的酶切谱系并进行聚类分析，尝试将该方法用于人类致病相关毛孢子菌的快速种间鉴定。方法 收集8种14株毛孢子菌，对其进行rDNA-ITS/IGS1扩增测序，并分别应用HaeⅢ、HhaⅠ、HaeⅢ和HhaⅠ双酶、HinfⅠ、MspⅠ和TaqⅠ对扩增片段进行RFLP酶切。结果 rDNA-ITS-RFLP多酶切可将8种毛孢子菌分为不同进化分支的4个亚类，而rDNA-IGS1-RFLP多酶切则可实现8种14株毛孢子菌准确的种间鉴定，并实现对部分进化距离较远的阿萨希毛孢子菌基因型之间的区分。结论 rDNA-ITS/IGS1-RFLP多酶切技术有望用于毛孢子菌属种水平的快速鉴定。

关键词: 毛孢子菌, 种间鉴定, 核糖体DNA, PCR-RFLP

Abstract:

Objective To genotype Trichosporon spp. with rDNA-ITS/IGS1-RFLP analysis followed by cluster analysis, and attempt to apply this method to rapid species identification of human pathogenic Trichosporon spp.. Methods Fourteen strains of Trichosporon, which belonged to 8 species, were collected. The rDNA-ITS/IGS1 regions were amplified by PCR and sequenced. Simultaneously, the amplicons were digested separately with restriction enzymes, including HaeⅢ, HhaⅠ, HaeⅢ and HhaⅠ, HinfⅠ, MspⅠ and TaqⅠ. Results The 8 species of Trichosporon could be classified into 4 subgroups with rDNA-ITS-RFLP, while interspecies identification of all the 14 strains from 8 species of Trichosporon could be realized with rDNA-IGS1-RFLP. Also, those genotypes of T. asahii which had relative long phylogenic distance could even be discriminated with rDNA-IGS1-RFLP. Conclusion The rDNA-ITS/IGS1-RFLP analysis is expected to be used in rapid interspecific identification of genus Trichosporon.

Key words: Trichosporon, Interspecific identification, ribosome DNA, PCR-RFLP